GSM6541883: siCtrl G9a ChIP-seq Rep1; Homo sapiens; ChIP-Seq
Sample information curated by ChIP-Atlas
Antigen
Antigen Class
TFs and others
Antigen
EHMT2
Cell type
Cell type Class
Prostate
Cell type
LNCAP
Primary Tissue
Prostate
Tissue Diagnosis
Carcinoma
Attributes by original data submitter
Sample
source_name
Human prostate cancer cells
cell line
LNCaP
treatment/infection
siCtrl
antibody
G9a
vendor
bethyl laboratory
catalog number
A300-933A
lot/batch number
Lot 2
Sequenced DNA Library
library_name
GSM6541883
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
For SUZ12 and G9a ChIP-Seq, Chromatin in nuclear fraction were sheared to 200-500bp using an Covaris M220 Focused-ultrasonicator, Lysates were clarified from sonicated nuclei and protein-DNA complexes were immunoprecipitated using the indicated antibody. DNA was then reverse crosslinked from protein and purified. For H3K27me3, H3K9me2 CUT&Tag and PALI1 CUT&RUN, 0.5x10^6 LNCaP cells bound to Concanavalin A-coated beads were incubated with indicated primary antibodies and secondary antibody. pA-Tn5 was used to induce DNA tagmentation in CUT&Tag while pA-MNase was used to digest chromatin/DNA bound by the antibodies-protein complex in CUT&RUN, followed by Proteinase K digestion and PCI extraction. For CUT&Tag, the ethanol precipitated DNA was immediately subjected to PCR amplification by Universal and barcoded i7 primer using NEBNext HiFi 2x PCR Master mix. The amplified library was purified by using Agencourt AMPure XP beads without size selection. For ChIP-Seq and CUT&RUN, ChIP-seq libraries were prepared from 3-5ng ChIPped DNA while CUT&RUN libraries were prepared from 15-20ng DNA using NEBNext® Ultra™ II DNA Library Prep Kit (NEB, E7645S), according to the manufacturer's instructions. Postamplification libraries were size selected at 250–450 bp (ChIP-Seq) and 160-320bp (CUT&RUN) in length using Agencourt AMPure XP beads from Beckman Coulter and were quantified using the Library Quantification Kit from Illiumina (Kapa Biosystems, KK4603). All CUT&Tag, CUT&RUN and ChIP-Seq Libraries were pooled to a final concentration of 10nM and sequenced single-end using the Illumina HiSeq 4000.